THE ROLE OF MECHANICALLY GATED ION CHANNELS IN REGULATING THE RESPONSE OF MESENCHYMAL STEM CELLS TO NANOFIBER ARCHITECTURE

Open Access
Author:
Wandling, Grant Donald
Area of Honors:
Biomedical Engineering
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
  • Justin Lee Brown, Thesis Supervisor
  • William O Hancock, Honors Advisor
Keywords:
  • Nanofibers
  • TREK-1
  • hMSCs
  • osteoblasts
Abstract:
The human body is a constantly changing, heterogeneous structure often in need of assistance in times of trauma and aging. Over one hundred billion dollars are spent on orthopedic trauma, malignancy, infection, and degradation care and repair. Many of these cases require intervention to achieve and maintain union between healing bone components, which is needed by bone cells to properly correct the injured site. The living cells within the human body must perceive and react to their surroundings in order to carry out their basic functions and repair damaged tissues. The physical interaction between a cell and the extracellular matrix is a complex structure and process comprised of many protein interactions. A cell uses these interactions to interpret and respond to the changes in their surroundings, often by converting a physical force into an electrochemical signal. These signals are sometimes created by the movement of ions across a membrane, through a transmembrane channel. TREK-1 is a transmembrane potassium channel responsible for maintaining the cell membrane potential. We hypothesize that changes in the extracellular architecture will alter the expression of TREK-1. Alterations of the extracellular geometry will lead to changes in the forces applied to cells, eliciting an electrochemical response, which must be regulated by ion channels such as TREK-1. By using techniques such as western blotting, ELISA, and immunofluorescence imaging, we can detect concentrations of TREK-1 in osteoblast and human mesenchymal stem cells. While no trend was observed due to low protein concentrations, the presence of TREK-1 was verified in multiple cell lines. Future work in this area will look to expand the variety of extracellular architectures and better quantify protein expression levels on these substrates.